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In Vitro Cellular & Developmental Biology. Animal

Gulbahar Boyuk, A Arzu Yigit, Ilkay Aydogan
Islet cell transplantation is a major treatment strategy for type I diabetes, and has proven to be effective for maintaining glucose homeostasis. However, this treatment requires an extended period of immunosuppression to prevent rejection and recurrent transplantation to maintain function. Thus, to enhance the properties of transplanted islet cells, we examined the effect of the co-culture of luteal cells, which secrete progesterone, on islet cell viability, functionality, and revascularization. It was found that islet viability and functionality were higher in the co-cultured group than in single cultures of islets at 48 and 96 h, in parallel with increased progesterone and vascular endothelial growth factor (VEGF) secretion from luteal cells...
September 5, 2018: In Vitro Cellular & Developmental Biology. Animal
Azim Parandakh, Mohammad Tafazzoli-Shadpour, Abdolreza Ardeshirylajimi, Arash Khojasteh, Mohammad-Mehdi Khani
Cellular mechanical characteristics represent cell ability to produce tissue-specific metabolites. Therefore, to achieve effective cell therapy, a better understanding of the effects of chemo-mechanical stimuli on the mechanical properties of in vitro-treated cells is essential. Herein, we investigated the effects of uniaxial strain on the mechanical properties of mesenchymal stem cells (MSCs) upon transforming growth factor beta 1 (TGF-β1 ) stimulation. The MSCs were categorized into control and test groups...
September 3, 2018: In Vitro Cellular & Developmental Biology. Animal
Payal Agarwal, Patricia DeInnocentes, R Curtis Bird
p16 is an important tumor suppressor gene encoded by the INK4A/ARF/INK4B gene locus that is conserved in humans, rodents, and canids. p16 regulates cell cycle in early G1 phase inhibiting transition out of cell cycle from G1/S phase by regulating a multi-protein control complex. p16-associated proteins, cyclin D, CDK4, and CDK6, experience expression level decreases or do not change during cell differentiation and quiescence in contrast to constant p16 expression in post-proliferative cell phases. We hypothesized that p16 has alternate binding partners, other than classical proliferation-associated proteins such as CDKs, in these post-proliferative cell phases...
August 30, 2018: In Vitro Cellular & Developmental Biology. Animal
Masamichi Hayashi, Naoki Yamamoto, Noriko Hiramatsu, Sumito Isogai, Yusuke Gotoh, Yasuhiro Goto, Masashi Kondo, Kazuyoshi Imaizumi
In recent research on regenerative medicine, three-dimensional (3D) tissue reconstruction using the induced pluripotent stem cell (iPS cell) differentiated cells has attracted attention. In this study, mouse lungs at 1.5, 10, and 20 d old were subjected to enzyme treatment, and aggregates formed in serum-free suspension culture (3D-culture) were observed. The number of aggregates formed was the highest in 1.5 d. The cell aggregates in which the interior of the aggregate is filled and form small vacuoles and the organoid-like aggregates having a relatively large vacuole inside and forming the alveolar-like structure were observed...
August 25, 2018: In Vitro Cellular & Developmental Biology. Animal
Jirapat Namkaew, Thiranut Jaroonwitchawan, Narawadee Rujanapun, Jantip Saelee, Parinya Noisa
Neuroblastoma is the most common cancer of the sympathetic nervous system in children. Here, the influences of curcumin on survival, apoptosis, migration, and its combined effects with doxorubicin were investigated in SH-SY5Y cells by cell survival assay, flow cytometry, migration assays, and RT-PCR. Curcumin inhibited SH-SY5Y cell growth and induced apoptosis in dose- and time-dependent manners. This apoptotic induction relied on the upregulation of p53 and p21. Moreover, the treatment of curcumin for 24 h significantly suppressed cell migration, together with the downregulation of matrix metalloproteinase-2 (MMP-2) and upregulation of tissue inhibitor of metalloproteinases-1 (TIMP-1)...
August 22, 2018: In Vitro Cellular & Developmental Biology. Animal
Jun Zhang, Lei Xiong, Wenxian Tang, Lin Tang, Baohe Wang
This study aimed to examine the proliferative behavior and molecular mechanisms of rat bone marrow-derived MSCs (rBMSCs) cultured under three different oxygen concentrations. Passaged rBMSCs exhibited significantly greater proliferation rates at 1% O2 and 5% O2 than those at 18% O2 and the cells exposed to 1% O2 showed the highest proliferative potential, which was evidenced by the growth curves, colony-forming efficiencies, and CCK-8 absorbance values. The rBMSCs grown under hypoxic culture conditions (1% O2 and 5% O2 ) had the increased percentage of cells in S + G2 /M-phase and the decreased apoptotic index, compared with normoxia (18% O2 )...
August 22, 2018: In Vitro Cellular & Developmental Biology. Animal
Vahid Kia, Maryam Sharif Beigli, Vahedeh Hosseini, Ameneh Koochaki, Mahdi Paryan, Samira Mohammadi-Yeganeh
Breast cancer is the first common cancer among women worldwide. One of the major signaling pathways playing a role in the onset and progression of this disease is PI3K/Akt/mTOR, which can be inhibited by PTEN. miRNAs are small non-coding molecules that regulate the expression of their targets by inhibition or suppression, and thus, their dysregulated expression results in the development of cancer. Using various software applications predicting miRNAs and evaluating GEO microarray data, miR-144 was selected as an inhibitor of PTEN...
August 21, 2018: In Vitro Cellular & Developmental Biology. Animal
Hao Xu, Na Wang, Wenyue Cao, Liang Huang, Jianfeng Zhou, Lingshuang Sheng
Nowadays, adoptive T cell immunotherapy is emerging as a novel and potent treatment for cancer. To prepare enough effective T cells for treatment use, their rapid expansion is favorable. Our study compared 6 commonly used cultural media for human T cells, including serum-containing media and serum-free media, namely RPMI 1640, IMDM, Gibco OpTmizer CTS T Cell Expansion SFM, Gibco AIM-V Medium CTS, LONZA X-VIVO 15, and StemSpan SFEM with or without Dynabeads Human T-Activator CD3/CD28, on in vitro T cell expansion, apoptosis, and immune phenotype...
July 12, 2018: In Vitro Cellular & Developmental Biology. Animal
Joanne Keenan, Karina Horgan, Martin Clynes, Indre Sinkunaite, Patrick Ward, Richard Murphy, Finbarr O'Sullivan
Copper is an essential trace element micronutrient in human and animal nutrition. Trace amounts present even in ultrapure water, serum and other cell culture medium components are sufficient to support the health requirements of most cell types in culture. Analysis of a variety of different types of basal media from a number of different suppliers revealed large fluctuations in the levels of copper, and also of other micronutrients including zinc, iron, selenium and cobalt. Investigations on proliferating Caco-2 cells revealed reductions in growth with increasing copper concentrations within the range seen in the commercial media and changes in expression of apoptosis- and autophagy-related proteins were noted...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Rongqiang Wu, Peng Xue, Yu Wan, Shizhong Wang, Meng Gu
Uc.40 is a long noncoding RNA that is highly conserved among different species, although its function is unknown. It is highly expressed in abnormal human embryonic heart. We previously reported that overexpression of uc.40 promoted apoptosis and inhibited proliferation of P19 cells, and downregulated PBX1, which was identified as a potential target gene of uc.40. The current study evaluated the effects of uc40-siRNA-44 (siRNA against uc.40) on the differentiation, proliferation, apoptosis, and mitochondrial function in P19 cells, and investigated the relationship between uc...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Change Gao, Qian Song, Ming Zhang, Jian Li, Miao Yi, Jian Dong
T cells have been successfully applied to cancer immunotherapy. However, a challenge is an expansion of T cells from cynomolgus monkey, which is a valuable non-human primate model for T cell therapy transferring to the clinic. Here, we compared several strategies to expand cynomolgus monkey T cell and developed an appropriate method. Our study demonstrated that 100 ng/ml CD3 mAb + 1% PHA+ 1000 U/ml IL2 therapy significantly expanded peripheral blood CD3+ T cells without compromising T cell phenotype in vitro...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Liping Lu, Haicheng Zhang, Weifeng Dong, Wenjia Peng, Jing Yang
The mechanisms for cardiomyocyte death in cardiovascular diseases are incompletely understood. The aim of this study is to reveal the function of miR-381 in myocardium infarction (MI)-induced cardiomyocyte apoptosis. We established mouse model of MI and cellular models of apoptosis induced by oxidative stress (H2 O2 and hypoxia/reoxygenation (H/R)). The expression of miR-381 in these models was assessed by quantitative reverse transcription polymerase chain reaction (qRT-PCR); we employed approaches including cell counting kit-8 (CCK-8) assay and flow cytometry to evaluate the cell viability and apoptosis...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Rong-Shing Chang, Chi-Shuo Chen, Ching-Lung Huang, Chiu-Ting Chang, Yujia Cui, Wei-Ju Chung, Wun-Yi Shu, Chi-Shiun Chiang, Chun-Yu Chuang, Ian C Hsu
Application of high-dosage UVB irradiation in phototherapeutic dermatological treatments present health concerns attributed to UV-exposure. In assessing UV-induced photobiological damage, we investigated dose-dependent effects of UVB irradiation on human keratinocyte cells (HaCaT). Our study implemented survival and apoptosis assays and revealed an unexpected dose response wherein higher UVB-dosage induced higher viability. Established inhibitors, such as AKT- (LY294002), PKC- (Gö6976, and Rottlerin), ERK- (PD98059), P38 MAPK- (SB203580), and JNK- (SP600125), were assessed to investigate UV-induced apoptotic pathways...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Min Chen, Hang Jie, Zhongxian Xu, Tianyuan Ma, Meiyan Lei, Dejun Zeng, Guijun Zhao, Xiaolan Feng, Chengli Zheng, Chenglu Zhang, Zhengjie Liang, Diyan Li
Research of epithelial cells in musk gland is lacking. There are no good characterized epithelial cell lines that can provide complementary in vitro models for in vivo research. We successfully cultivated epithelial cells of musk gland for the first time. The protocol described here produces epithelial cell lines from the mature secreting musk gland. Based on morphological observation, epithelial cells of musk gland were isolated and cultured in vitro. After the third passage, the musk gland-derived cells were filled with many lipid droplets and proliferated well...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Ann M Fallon
Wolbachia pipientis (Rickettsiales; Anaplasmataceae) is an obligate intracellular alpha proteobacterium that occurs in arthropods and filarial worms. Some strains of Wolbachia can be maintained as persistent infections in insect cell lines. C/wStr1 cells from the mosquito Aedes albopictus maintain a robust infection with Wolbachia strain wStr, originally isolated from the planthopper, Laodelphax striatellus. To explore possible functions of penicillin-binding proteins expressed from the wStr genome, C/wStr1 cells were exposed to ampicillin...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Isha Verma, Polani B Seshagiri
P19 embryonal carcinoma cells (EC-cells) provide a simple and robust culture system for studying neural development. Most protocols developed so far for directing neural differentiation of P19 cells depend on the use of culture medium supplemented with retinoic acid (RA) and serum, which has an undefined composition. Hence, such protocols are not suitable for many molecular studies. In this study, we achieved neural differentiation of P19 cells in a serum- and RA-free culture medium by employing the knockout serum replacement (KSR) supplement...
September 2018: In Vitro Cellular & Developmental Biology. Animal
Minh Nguyen Tuyet Le, Mika Takahi, Kenshiro Maruyama, Akira Kurisaki, Kiyoshi Ohnuma
A high density of human-induced pluripotent stem cells (hiPSCs) improves the efficiency of cardiac differentiation, suggesting the existence of indispensable cell-cell interaction signals. The complexity of interactions among cells at high density hinders the understanding of the roles of cell signals. In this study, we determined the minimum cell density that can initiate differentiation to facilitate cell-cell interaction studies. First, we co-induced cardiac differentiation in the presence of the glycogen synthase kinase-3β inhibitor CHIR99021 and activin A at various cell densities...
August 2018: In Vitro Cellular & Developmental Biology. Animal
Vidya Jayasankar, Srinivasa Raghavan Vasudevan, Suja C Poulose, Indira Divipala
Mantle tissue from the black-lip pearl oyster, Pinctada margaritifera, was cultured in vitro using sterilized seawater supplemented with 0.1% yeast extract as the culture medium. Granular and agranular epithelial cells, hyalinocytes, and fibroblast-like cells were observed in the initial stages of culture. Epithelial cells later formed pseudopodial cell networks containing clusters of granulated cells, which upon maturation released their colored granules. These granules induced formation of nacre crystal deposits on the bottom of the culture plate...
August 2018: In Vitro Cellular & Developmental Biology. Animal
Michihiro Toritsuka, Manabu Makinodan, Takahira Yamauchi, Yasunori Yamashita, Daisuke Ikawa, Takashi Komori, Sohei Kimoto, Kaori Hamano-Iwasa, Hideo Matsuzaki, Toshifumi Kishimoto
Lymphoblastoid cell lines (LCLs) are nearly immortalized B lymphocytes that are used as long-lasting supply of human cells for studies on gene expression analyses. However, studies on the stability of the cellular features of LCLs are scarce. To address this issue, we measured gene expression in LCLs with different passage numbers and observed that gene expression substantially changed within 10 passages. In particular, the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a well-known housekeeping gene, varied considerably during subculture; thus, the use GAPDH as an internal control may be unsuitable...
August 2018: In Vitro Cellular & Developmental Biology. Animal
Fusako Kito, Rieko Oyama, Marimu Sakumoto, Mami Takahashi, Kumiko Shiozawa, Zhiwei Qiao, Hiromi Sakamoto, Takeshi Hirose, Nokitaka Setsu, Akihiko Yoshida, Akira Kawai, Tadashi Kondo
Osteosarcoma is an aggressive mesenchymal malignancy of the bone. Patient-derived models are essential tools for elucidating the molecular mechanisms associated with poor prognosis and the development of novel anticancer drugs. This study described the establishment of a patient-derived cancer model of osteosarcoma. Primary osteosarcoma tumor tissues were obtained from an osteosarcoma patient and inoculated in the skin of immunodeficient mice, followed by transplantation to other mice upon growth. Cells were maintained in monolayer cultures, and the capability of spheroid formation was assessed by seeding the cells on culture dishes...
August 2018: In Vitro Cellular & Developmental Biology. Animal
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